Studies conducted in the course of this grant are aimed at establishing the significance of PCR based assays for the diagnosis of pediatric HIV-1 infection and prediction of eventual progression to disease. A simple, rapid, reliable assay suitable for routine clinical testing will be developed and evaluated for the direct detection of HIV-1 sequences in seropositive babies. This assay will consist of polymerase chain reaction (PCR) amplification of two regions of the HIV-1 genome coupled with an oligomer capture assay in a microtiter well format. The specificity and sensitivity of this assay will be evaluated on laboratory control specimens, and an optimum format selected. The procedures for complete PCR testing of clinical samples including sample preparation, amplification, and detection will be streamlined to reduce handling and incubation times. Appropriate controls will be provided to validate the results of the assay. Additional modifications to the assay will be designed to permit detection of HIV-1 RNA sequences and to quantitate the PCR results. The RNA test may serve as a useful indicator of active infection and may be predictive of progression of disease. The quantitative assay will permit measurement of viral load and be useful for monitoring the response to anti-retroviral treatment. The tests developed in the course of this grant will permit more meaningful diagnosis of HIV-1 infected babies and allow better management of infection.